On 14 May 1997 11:27:39 +0100, Claus Rodemer
<c.rodemer at dkfz-heidelberg.de> wrote:
>Hi all,
>which is the best way to make blunt ends ? I have cut the DNA with a enzym
>that makes 3' protruding ends and I want to make blunt ends out of it.
>>Is Klenow Ok, or T4 DNA-Pol ? Or is it possible with S1 Nuclease,too?
>>Another thing:
>I tested a cloning kit, and with this kit there is a "polishing reaction"
>provided (cloned Pfu Pol) .The manual said that this makes blunt ends ,too.
>Well I am still testing ... anybody knows something about this ?
>>Bye,
>Claus
>--
>>>------------------------------------------------------------------------
>Claus Rodemer German Cancer Research Center INF 280
>Div.Biophysics of Macromolecules AG. Gene Regulation and DNA Topology
>D-69120 Heidelberg E-Mail: C.Rodemer at dkfz-heidelberg.de>WWW:http://www.rzuser.uni-heidelberg.de/~crodemer/>------------------------------------------------------------------------
>>S1 nuclease should not be used because there is a tendency to loose
also base-paired nucleotides, when overdigestion is accidentally
performed, I always use Klenow with good success.
Martin